The cap‐snatching reaction of yeast L‐A double‐stranded RNA virus is reversible and the catalytic sites on both Gag and the Gag domain of Gag‐Pol are active

Summary The yeast L‐A double‐stranded RNA virus synthesizes capped transcripts by a unique cap‐snatching mechanism in which the m 7 Gp moiety of host mRNA (donor) is transferred to the diphosphorylated 5′ end of the viral transcript (acceptor). This reaction is activated by viral transcription. Here, we show that cap snatching can be reversible. Because only m 7 Gp is transferred during the reaction, the resulting decapped donor, as expected, retained diphosphates at the 5′ end. We also found that the 5′ terminal nucleotide of the acceptor needs to be G but not A. Interestingly, the A‐initiated molecule when equipped with a cap structure (m 7 GpppA…) could work as cap donor. Because the majority of host mRNAs in yeast have A after the cap structures at the 5′ ends, this finding implies that cap‐snatching in vivo is virtually a one‐way reaction, in favor of furnishing the viral transcript with a cap. The cap‐snatching sites are located on the coat protein Gag and also the Gag domain of Gag‐Pol. Here, we demonstrate that both sites are functional, indicating that activation of cap snatching by transcription is not transmitted through the peptide bonding between the Gag and Pol domains of Gag‐Pol.